Bovines are the main intermediary host of the protozoan Neospora caninum, which is a major cause of bovine abortions and neonatal mortality worldwide. Sera were collected from 373 dairy cattle and 33 dogs on 19 dairy farms in Lages city, Santa Catarina State, Brazil, to determine the prevalence of N. caninum antibodies and risk factors. Tests for N. caninum antibodies were done using an indirect immunofluorescent antibody test (IFAT). Positive reactions with titers ≥ 1:200 (cattle) and ≥ 1:50 (dogs) were found in 86 (23.1%) and seven (21.2%) bovines and dogs, respectively. Of the bovines, four (1.1%) were calves, 33 (8.8%) were heifers, 314 (84.2%) were lactating cows, 19 (5.1%) were dry cows and three (0.8%) were bulls. Infection was primarily found in animals with low milk production (P = 0.008). The presence of seropositive dogs on the properties did not affect the prevalence of bovine infection. No correlation between bovine reproductive disorders and the prevalence of N. caninum antibodies was observed. Our results showed that N. caninum infection is widespread among dairy cattle of Lages, Santa Catarina.